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Calcium channels activated by hydrogen peroxide mediate abscisic acid signalling in guard cells


Drought is a major threat to agricultural production. Plants synthesize the hormone abscisic acid (ABA) in response to drought, triggering a signalling cascade in guard cells that results in stomatal closure, thus reducing water loss1. ABA triggers an increase in cytosolic calcium in guard cells ([Ca2+]cyt)2,3,4,5,6 that has been proposed to include Ca2+ influx across the plasma membrane3,5,7,8,9. However, direct recordings of Ca2+ currents have been limited3 and the upstream activation mechanisms of plasma membrane Ca2+ channels remain unknown. Here we report activation of Ca2+-permeable channels in the plasma membrane of Arabidopsis guard cells by hydrogen peroxide. The H2O2-activated Ca2+ channels mediate both influx of Ca2+ in protoplasts and increases in [Ca2+]cyt in intact guard cells. ABA induces the production of H2O2 in guard cells. If H2O2 production is blocked, ABA-induced closure of stomata is inhibited. Moreover, activation of Ca2+ channels by H2O2 and ABA- and H2O2-induced stomatal closing are disrupted in the recessive ABA-insensitive mutant gca2. These data indicate that ABA-induced H2O2 production and the H2O2-activated Ca2+ channels are important mechanisms for ABA-induced stomatal closing.

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Figure 1: H2O2 activates inward currents in Arabidopsis guard cells.
Figure 2: H2O2-activated currents are Ca2+-permeable, non-selective cation currents.
Figure 3: H2O2-activated Ca2+ currents mediate Ca2+ influx in guard cell protoplasts and H2O2-induced [Ca2+]cyt increases in intact guard cells.
Figure 4: ABA causes H2O2 production and H2O2-induced stomatal closing requires external Ca2+.
Figure 5: The recessive ABA-insensitive Arabidopsis mutant gca2 disrupts H2O2 activation of Ca2+ currents and ABA-induced stomatal closing.

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We thank J. Satimanont and C. Kwak for assistance; R. Tsien and M. Yanofsky for use of imaging systems and fluorescence microscopes; and J. Kwak, D. Lee, R. Vaidyanathan, M. Ghassemian, N. Leonhardt and P. Buschmann for comments on the manuscript. This work was supported by Department of Energy, National Institutes of Health and National Science Foundation grants to J.I.S. and by a European Biotech Program grant and Fonds der Chemischen Industrie to E.G. Y.M was supported by a fellowship for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, S.T and G.A by Human Frontier Science Program fellowships, and B.K. by a Feodor-Lynen fellowship from the Alexander von Humboldt Foundation.

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Correspondence to Julian I. Schroeder.

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Pei, ZM., Murata, Y., Benning, G. et al. Calcium channels activated by hydrogen peroxide mediate abscisic acid signalling in guard cells. Nature 406, 731–734 (2000).

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